辽宁绒山羊CSN3基因PCR-RFLP多态性分析

本试验以辽宁绒山羊和内蒙古绒山羊为实验动物,采用PCR-RFLP技术对-酪蛋白基因(CSN3)片段的Ⅰ的酶切多态性进行分析,结果表明:辽宁绒山羊和内蒙古绒山羊2个山羊群体中均存在CSN3基因Ⅰ酶切位点的多态性,且均有A、B两个等位基因。辽宁绒山羊群体等位基因A和B的基因频率分别为0.46和0.54;内蒙古绒山羊群体中等位基因A和B的基因频率分别为0.31和0.69。CSN3基因Ⅰ酶切位点的基因型分布在辽宁绒山羊群体中极显著偏离Hardy-Weinberg平衡定理(<0.01),在内蒙古绒山羊群体中符合Hardy-Weinberg平衡定理(>0.05),但两个群体间存在显著差异(<0.01)。     

Effect of nitric oxide on urotensin-Ⅱ expression in pulmonary arterioles of rats chronically exposed to hypoxia-hypercapnia

AIM: To investigate the roles of nitric oxide/L-arginine (NO/L-Arg) pathway and urotensin-Ⅱ (UⅡ) in the development of pulmonary hypertension induced by chronic hypoxia-hypercapnia in rats.METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups (n=10): normal control group (A), hypoxia-hypercapnia+saline group (B), hypoxia-hypercapnia+L-Arg liposome group (C) and hypoxia-hypercapnia+N-nitro-L-arginine methyl ester (L-NAME) group (D).Contents of UⅡ, UⅡ mRNA and receptor of UⅡ (UT) mRNA in pulmonary arterioles were measured with immunohistochemistry analysis and in situ hybridization, respectively.Change of small pulmonary vascular microstructure was also investigated.RESULTS: (1) The mean pulmonary artery pressure (mPAP) and the weight ratio of right ventricle to left ventricle plus septum ［RV/(LV+S)］ in B and D groups were all higher than those in A group (respectively, P<0.05), with C group significantly lower than those in B group (respectively, P<0.01).(2) Light microscopy showed that the ratio of vessel wall area to total area (WA/TA) and the media thickness of pulmonary arterioles (PAMT) in B group were higher than those in A group (P<0.05), with C group significantly lower than those in B group.(3) The contents of UⅡ, UⅡ mRNA and UT mRNA in pulmonary arterioles in B and D groups were all higher than those in A group (respectively, P<0.01), while the expression of UⅡ and UⅡ mRNA in C group were lower than those in B group (P<0.01).CONCLUSION: The pathological process of pulmonary hypertension induced by chronic hypoxia-hypercapnia might be related to upregulation of UⅡ located in pulmonary arterioles, which might be partially inhibited by exogenous NO in rats.     

HP-PRRSV抑制猪肺eNOS表达与猪肺微血栓形成的相关性研究

为了解高致病性猪繁殖与呼吸综合征病毒（HP-PRRSV）与肺微血栓病变的关系及内皮型一氧化氮合酶（eNOS）的表达对微血栓的影响,选取50头70日龄的鄂通两头乌猪,4头作为正常对照,46头感染HP-PRRSV。采取50头猪的肺组织,应用HE染色观察组织病理变化并对微血栓病变程度进行等级评分,将46头感染猪分为轻度微血栓组、中度微血栓组、重度微血栓组,每组选取4头猪进行后续研究。应用免疫组化、荧光定量PCR和Western blot技术,明确HP-PRRSV和eNOS在微血栓猪肺中的表达、分布及变化。组织病理学研究结果显示感染猪肺组织肺泡间隔明显增宽,毛细血管有不同程度的淤血、出血,肺泡腔内有巨噬细胞、脱落的肺泡上皮细胞及淋巴细胞,肺泡壁有大量微血栓。HP-PRRSV主要分布于肺泡巨噬细胞及少量的淋巴细胞的细胞质。随着HP-PRRSV病毒含量的增加,微血栓病变越严重。微血栓病变越严重,死亡率越高。eNOS主要分布于肺泡Ⅱ型细胞、巨噬细胞、血管内皮细胞及支气管平滑肌细胞的细胞质,感染猪肺组织的eNOS mRNA和蛋白表达显著低于未感染对照组。研究结果表明HP-PRRSV使eNOS的表达下调,并参与感染猪肺组织微血栓的形成,增加感染猪的死亡率。     

不同杀菌剂及复配对苜蓿锈病的防治研究

为了筛选防治苜蓿(Medicago sativa)锈病有效单剂和复配组合,本试验采用盆栽喷雾法,测定了14种杀菌剂及5种复配剂对苜蓿锈菌的毒力,并比较不同复配的协同增效作用。结果表明:在单剂毒力测定中,苜蓿锈菌对嘧菌酯最敏感,EC₅₀为1.8809mg·L^-1;甲基硫菌灵的毒力最弱,EC₅₀为747.0022mg·L^-1。按照不同杀菌机理的药剂复配机理,嘧菌酯和戊唑醇复配中,1:9组合产生相加作用,其余比例复配均表现为增效作用,其中5:1增效作用最强,增效系数SR为4.9684;嘧菌酯和烯酰吗啉复配,协同增效作用最强的是以1:3组合,增效系数为5.1114,但二者以9:1复配时产生拮抗作用;嘧菌酯和代森锰锌复配,1:7复配组合增效系数SR为4.5757,协同增效作用比较明显。吡唑醚菌酯与苯醚甲环唑复配增效作用最强的是1:5复配组合,其增效系数SR为5.2276;与百菌清的复配,1:7复配组合增效作用最为明显,其增效系数SR为5.8058,因此,从单剂防效看嘧菌酯和吡唑醚菌酯效果最好,复配剂中,吡唑醚菌酯和百菌清以1:7防效最好。     

鸭瘟诊断方法研究进展

鸭瘟是由疱疹病毒引起的鸭、鹅的一种烈性传染病。目前已有多种诊断方法用于鸭瘟的诊断。本文就鸭瘟诊断方法的研究进展作一综述。     

鸽禽Ⅰ型副粘病毒病蜂胶佐剂灭活疫苗的研制

鸽禽Ⅰ型副粘病毒（A/PMV-Ⅰ）病是鸽的一种严重病毒性传染病，给养鸽业造成了巨大损失。为了更好地预防本冱的发生，我们用从现场分离鉴定的7株鸽A/PMV-Ⅰ毒株中筛选出3株抗原性良好的毒株作为种毒，研制成蜂胶佐剂混合毒株灭活疫苗，经安全试验、最佳免疫剂量测定、铲力试验，以及田间应用的结果，表明该疫苗抗原性良好、使用安全和免疫保护效果确定。实验室试验，用本疫苗免疫后30、90、180天，其保护脂数分别为100%、77.8%和55.6%,免疫有效期最少达6个月；在4℃-8℃保存,其有效其最少可达180天。     

溴尿嘧啶处理受精卵对中国对虾抗病和生长的影响

以5-溴尿嘧啶(5-Bromouracil,5-BrUra)为诱变剂,分别以前期实验在半致死条件下获得的处理浓度、时间组合0.5 mg/ml 9 h(0.5～9)、0.3 mg/ml 16 h(0.3～16)来处理受精卵,培养获得诱变材料,对处理群体抗病力和生长性状进行分析,探讨化学诱变剂对中国对虾Fenneropenaeus chinensis抗白斑综合征病毒(White Spot Syndrome Virus,WSSV)能力和生长的影响.WSSV攻毒实验结果表明,0.5～9处理组、0.3～16处理组和对照组存活时间分别为77.20±3.71 h、74.57±2.88 h和81.45±2.98 h,三者之间不存在显著差异,说明在本实验条件下,5-溴尿嘧啶处理受精卵不能显著提高中国对虾对WSSV的抵抗能力(P>0.05).生长实验结果表明,在实验开始第21天至第60天之间,两处理组特定生长率(SGR)显著高于对照组;实验结束时,经过5-溴尿嘧啶处理的中国对虾群体的体长、体重日增长率显著高于对照组,且两个处理组之间没有显著差异,表明经5-溴尿嘧啶处理可能促进中国对虾的生长.但这些生长特性能否稳定遗传给后代尚需进一步研究.     

Influence of paricalcitol on renal tubulointerstitial fibrosis in diabetic nephropathy

AIM: To investigate the effect of paricalcitol (P) on renal tubulointerstitial fibrosis and the underlying mechanisms in diabetic nephropathy (DN).METHODS: DN rat model was induced by a single intraperitoneal injection of streptozotocin after fasting. The animals were randomly divided into 2 groups:the DN rats in paricalcitol-intervened group (group P) were injected intraperitoneally with paricalcitol dissolved in propylene glycol after the day when the model was induced successfully at a dose of 0.4 μg/kg (3 times a week); the DN rats in DN group (group D) were given isopyknic propylene glycol. Normal control group (group C) was also set up. The samples of blood, urine and renal tissue were collected after intervention of paricalcitol for 12 weeks. The biochemical indexes were measured. The renal tissues were used for pathologic observation and determining the expression of transforming growth factor-β1 (TGF-β1), Wnt-4, β-catenin and Klotho by immunohistochemistry and Western blotting. In addition, the correlation among the above indexes was analyzed.RESULTS: (1) Scr, BUN and 24 h urine protein increased significantly in group D compared with group C, while decreased in group P compared with group D (P<0.05). (2) The area of renal tubulointerstitial fibrosis increased in group D compared with group C, while decreased in group P compared with group D (P<0.05). (3) The expression of Klotho decreased, while the expression of TGF-β1, Wnt-4 and β-catenin increased in group D compared with group C (P<0.05). Compared with group D, the expression of Klotho increased, while the expression of TGF-β1, Wnt-4 and β-catenin decreased in group P (P<0.05). (4) The expression of Klotho was negatively correlated with the fibrosis area, TGF-β1, Wnt-4 and β-catenin (P<0.05).CONCLUSION: Paricalcitol inhibits renal tubulointerstitial fibrosis in DN by promoting the expression of renal Klotho, and inhibiting Wnt/β-catenin signaling pathway activation and TGF-β1 synthesis.     

不同施肥方式对红壤蔬菜田氨氧化细菌和氨氧化古菌群落的影响

通过构建氨单加氧酶基因(amoA)克隆文库,研究在红壤蔬菜田上只施用磷钾化肥(PK)、只施氮磷钾化肥(NPK)、施用腐熟有机肥(DNPK)和施用新鲜有机肥(FNPK)等4种不同施肥处理的土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)群落多样性及与土壤脲酶活性的相关性。结果表明:施加有机肥处理(DNPK和FNPK)的蔬菜田土壤的AOB文库和AOA文库OTU数量和Shannon指数高于只施用无机肥(NPK和PK)处理的蔬菜田土壤;DNPK和FNPK处理的土壤优势AOB菌群为多形亚硝化叶菌(Nitrosolobus multiformis),比例分别为88.5%和68.5%,NPK和PK处理的土壤优势AOB菌群为亚硝化单胞菌属(Nitrosospirasp.),比例分别为54.8%和65.5%;DNPK、FNPK、NPK和PK处理土壤优势AOA菌群均为阿伯丁土壤亚硝化细杆菌侯选种(CandidatusNitrosotalea devanaterra),比例分别为90.9%、84.4%、77.8%和45.2%;施加有机肥处理(DNPK和FNPK)的土壤脲酶活性和氨氧化微生物的多样性指数都高于只施用无机肥处理(NPK和PK);AOA群落多样性指数与土壤脲酶活性呈显著正相关,而AOB群落多样性与土壤脲酶活性相关性不显著。总体来看,有机肥比无机肥处理提高了AOA和AOB群落多样性,且AOA在红壤蔬菜田土壤氨氧化过程中起着更为重要的作用。     

Study on Clinical Efficacy of Oligosaccharide Sulfate and Dextran Sulfate Against Bovine Mastitis

In order to detect the clinical efficacy of oligosaccharide sulfate(DEAE) and dextran sulfate (DS-4000) against bovine mastitis,56 cows with bovine mastitis were divided into 5 groups (DEAE group,DS-4000 group,DEAE＋penicillin group,DS-4000＋penicillin group and penicillin group),and their breasts were perfusion with different drugs twice one day for 3 days.The effective rates of drugs were observed by clinic symptoms.Milk samples were collected before and after treatment of drugs,and the bacterial negative conversion rate of each group were detected by microbial examination.The results showed that total effective rates of 5 groups were 10.00%,20.00%,58.33%,75.00% and 50.00%,respectively,and the bacterial negative conversion rates were 0,12.50%,50.00%,54.55% and 44.44%,respectively.In conclusion,the curative efficiencies of the polysaccharides against bovine mastitis were weak,but it could be improved by combination with penicillin.